JP5787361B2 - Chromatograph measurement method and chromatograph measurement apparatus - Google Patents

Description

  The present invention relates to a chromatographic measurement method and a chromatographic measurement apparatus for detecting a test substance using an insoluble carrier having a detection site capable of specifically fixing a test substance contained in a sample liquid.

  In recent years, a liquid (specimen liquid) containing a test material (specimen) that may contain a test substance is sent to an insoluble carrier, and the presence / absence of this test substance is determined using, for example, an immunoassay. Alternatively, a chromatographic measurement method has been developed for simple and rapid measurement of the amount. In the chromatographic measurement method, heavy equipment and equipment are not required, and a measurement result can be obtained simply by allowing the sample liquid to drop and allowing to stand for about 5 minutes to 10 minutes. Therefore, the chromatographic measurement method is widely used in many scenes, for example, clinical examinations in hospitals or laboratory tests in laboratories, as a simple and quick measurement method with high specificity.

  On the other hand, in clinical practice such as clinics, clinics, home medical care, etc., immunochromatograph measuring devices (immunochromatography readers) are used as POCT (Point of Care Testing) measuring devices for simple measurements regardless of clinical laboratory specialists. ) Is used. This immunochromatographic measuring apparatus measures the coloration amount of the reagent at the detection site of the loaded device, and enables highly sensitive and reliable measurement even for the coloration amount that is difficult to visually determine.

  For example, in Patent Document 1, in the chromatographic measurement method, in order to reduce a measurement error due to a difference in viscosity for each sample liquid, a development speed of the sample liquid to be developed in the insoluble carrier is obtained, and based on this development speed. It is disclosed that the time for starting the irradiation of the analysis light is set. In Patent Document 2, in order to quickly determine whether or not the amount of the dropped sample liquid is sufficient in the chromatographic measurement method, the development speed of the sample liquid to be developed in the insoluble carrier is obtained and obtained in advance. It is disclosed that the drop amount is estimated from the relationship between the developed speed and the drop amount of the sample liquid. Patent Document 3 discloses that in a chromatographic measurement method, in order to reduce noise due to a labeling substance in the background, a washing solution is developed after the sample solution is developed on an insoluble carrier.

JP 2006-162496 A JP 2009-85695 A JP 2009-216695 A

  However, the conventional method has a problem that even if the sample liquid has the same concentration, the measurement value varies for each measurement and it is difficult to perform quantitative measurement. This is because the development of the sample liquid in the insoluble carrier occurs due to the course, and the difference in each measurement of the amount of the sample liquid that actually passed through the detection site (total amount of the passed liquid) is large. If the total amount of liquid passing through can be made uniform for each measurement, the quantitativeness can be improved. However, the total amount of liquid passing through depends on various factors such as the presence or absence of coexisting substances in the sample liquid, the physical properties of the insoluble carrier, the humidity, and the dripping amount of the sample liquid. difficult.

  The present invention has been made in view of the above problems, and an object of the present invention is to provide a chromatographic measurement method and a chromatographic measurement apparatus that enable measurement with higher quantitativeness in chromatographic measurement.

In order to solve the above problems, the chromatographic measurement method according to the present invention is:
In a chromatographic measurement method for detecting a test substance using an insoluble carrier having a detection site capable of specifically fixing the test substance contained in the sample liquid,
Develop sample liquid in insoluble carrier,
Based on multiple images of the insoluble carrier acquired over time by the image acquisition means, calculate the development speed of the sample liquid and the amount of color at the detection site at a predetermined time after the arrival time when the sample liquid reaches the detection site And
Based on the development speed and the cross-sectional area of the insoluble carrier, calculate the amount of the sample liquid that actually passed through the detection site during the time from the arrival time to the predetermined time,
The coloration amount at the predetermined time is standardized based on the calculated amount of the sample liquid.

  In the chromatographic measurement method according to the present invention, the labeling substance is developed in the insoluble carrier together with the sample solution, and the development speed can be calculated based on the change in the position of the labeling substance represented in the plurality of images. preferable. In this case, the labeling substance can be disposed in a region upstream of the detection site, or can be premixed in the sample liquid.

  Alternatively, in the chromatographic measurement method according to the present invention, the development speed can be calculated based on a change in the position of the development front of the sample liquid.

  Further, in the chromatographic measurement method according to the present invention, it is preferable that the development speed is updated with time.

The chromatographic measurement apparatus according to the present invention is
In a chromatographic measurement apparatus for detecting a test substance using an insoluble carrier having a detection site capable of specifically fixing the test substance contained in the sample liquid,
An insoluble carrier;
Image acquisition means for acquiring a plurality of images of the insoluble carrier over time;
Based on the plurality of images, calculate the development speed of the sample liquid and the coloration amount of the detection site at a predetermined time after the arrival time when the sample liquid reaches the detection site. The amount of the sample liquid that actually passed through the detection site during the time from the arrival time to the predetermined time is calculated, and the coloration at the predetermined time is calculated based on the calculated amount of the sample liquid Computational processing means for normalizing the quantity is provided.

  In the chromatographic measurement apparatus according to the present invention, the calculation processing means is a labeling substance that develops in the insoluble carrier together with the sample liquid, and determines the development speed based on the change in the position of the labeling substance represented in the plurality of images. It is preferable to calculate. In this case, the chromatographic measurement apparatus may have a labeling substance disposed in a region upstream of the detection site.

  Alternatively, in the chromatographic measurement apparatus according to the present invention, the calculation processing means may calculate the development speed based on a change in the position of the development front of the sample liquid.

  In the chromatographic measurement apparatus according to the present invention, it is preferable that the calculation processing means updates the development speed with time.

  According to the chromatographic measurement method and the measurement apparatus of the present invention, the amount of the sample liquid that has actually passed through the detection site during the time from the arrival time to the predetermined time (total passage liquid amount) is calculated. Based on the passing liquid amount, the coloration amount at the predetermined time can be normalized. That is, the measured value of the coloration amount is corrected by the total passing liquid amount. As a result, in chromatographic measurement, even if the total amount of liquid passing through each measurement cannot be made uniform, measurement with higher quantitativeness becomes possible.

It is the schematic which shows the external appearance structure of the chromatograph measuring apparatus of this embodiment. It is the schematic which shows the internal structure of the chromatograph measuring apparatus of this embodiment. It is a schematic top view which shows the device for an assay loaded into a chromatograph measuring apparatus. It is a schematic bottom view which shows the device for an assay loaded into a chromatograph measuring apparatus. It is a schematic sectional drawing which shows the cross section of the device for an assay in the II-II line | wire of FIG. 3A. It is a schematic sectional drawing of an insoluble support | carrier. It is the schematic which shows the mode of the sample liquid expand | deployed in an insoluble support | carrier. It is the schematic which shows the workflow of the process of a measuring method. It is the schematic which shows the workflow of the other process of a measuring method. It is the schematic for demonstrating the other calculation method of expansion | deployment speed.

  Hereinafter, although an embodiment of the present invention is described using a drawing, the present invention is not limited to this. In addition, for easy visual recognition, the scale of each component in the drawings is appropriately changed from the actual one.

"Chromatograph measurement method and chromatograph measurement apparatus"
The chromatographic measurement method of the present embodiment is performed using, for example, the chromatographic measurement apparatus 1 shown in FIGS. 1 to 3. FIG. 1 is a schematic perspective view showing an external appearance of a chromatograph measuring apparatus 1 according to the present embodiment, and FIG. 2 is a schematic view showing the inside when an immunochromatographic device 20 is loaded and cut along the line II in FIG. FIG. 3A and 3B are a schematic top view and a schematic bottom view showing the appearance of the device 20, respectively, and FIG. 3C is a schematic cross-sectional view taken along the line II-II in FIG. 3A.

  Specifically, in the chromatographic measurement method of the present embodiment, there is a possibility that the test substance T is included in the device 20 having the test line T (detection site) and the control line C for determining the end of measurement. After supplying the sample liquid, the test apparatus is loaded with the device 20 in the measuring apparatus 1, developed the sample liquid in the insoluble carrier 21, and acquired by the image acquisition unit 40 over time (that is, a plurality of times along the time axis). Based on the plurality of images in the vicinity region 21a, the development speed of the sample liquid and the coloration amount of the test line T at a predetermined time after the arrival time when the sample liquid reaches the test line T are calculated, Based on the cross-sectional area of the insoluble carrier 21, the amount of the sample liquid that has actually passed through the test line T during the time from the arrival time to the predetermined time (total passage liquid amount) is calculated. It is intended to standardize the color former amount in the predetermined time based on the total flow-through volume.

  As shown in FIG. 1, the chromatographic measurement apparatus 1 according to the present embodiment includes a housing 10, a screen display unit 11 disposed on the surface of the housing 10, and an operation of a menu displayed on the screen display unit 11. A menu operation unit 12 and a power switch 13 for performing the above operation, and a device loading unit 14 for loading the device 20 into the apparatus. Further, as shown in FIG. 2, the chromatograph measurement apparatus 1 includes an image acquisition unit 40 that acquires an image of a region near the test line of the device 20, an information reading unit 50 that reads information from the device 20, and The memory 32 includes a calculation processing unit 34 that performs a predetermined calculation process based on the acquired image, and the image acquisition unit 40, the information reading unit 50, the memory 32, and a control unit 30 that controls the calculation processing unit 34. ing.

(Sample liquid)
The sample liquid that can be measured is particularly limited as long as it may contain a test substance (in addition to antigens, physiologically active substances such as natural products, toxins, hormones, and agricultural chemicals, or environmental pollutants). It is not something. For example, the sample fluid includes biological samples, particularly animal (especially human) body fluids (eg blood, serum, plasma, spinal fluid, tears, sweat, urine, pus, runny nose or sputum), excrement ( For example, feces), organs, tissues, mucous membranes and skin, an over-extracted specimen (swab) or gargle that is thought to contain them, or animals and plants themselves or their dried bodies diluted with a diluent, etc. it can.

  The sample solution is used as it is or in the form of an extract obtained by extracting the sample solution with an appropriate extraction solvent, and further, a diluted solution obtained by diluting the extract with an appropriate diluent. It can be used in the form or the extract concentrated by an appropriate method.

(Chromatography device)
3A, FIG. 3B, and FIG. 3C, the device 20 includes an insoluble carrier 21 having a test line T and a control line C, a device housing 22 for housing the insoluble carrier 21, A solution injection port 23 for injecting a reagent solution into the insoluble carrier 21 and an observation window 24 for observing a region 21a in the vicinity of the test line of the insoluble carrier 21 accommodated in the device housing 22 are provided. An information display unit 25 is provided on the surface of the housing 22.

  As shown in FIG. 4, the insoluble carrier 21 is soaked with a sample addition pad 81, which is a portion where the sample is dropped, and a labeling substance in the direction in which the sample such as the sample liquid is developed (direction of arrow A). A labeled substance holding pad 82, a chromatographic carrier 83 having a test line T (line-shaped detection site) on which a substance (specific binding substance) showing a specific binding property to a test substance is fixed, and a sent sample liquid And a transparent sheet 85 that supports the whole. Although one test line T is provided in the test line vicinity region 21a of the present embodiment, the number of test lines T is not limited to this and may be two or more. By using multiple test lines T and fixing different test substances, the presence and / or quantity of multiple test substances can be efficiently determined at a time when multiple test substances are included in the sample liquid. It becomes possible to measure. The material of the insoluble carrier 21 is preferably porous, for example, a nitrocellulose membrane, a cellulose membrane, an acetylcellulose membrane, a polysulfone membrane, a polyethersulfone membrane, a nylon membrane, glass fiber, a nonwoven fabric, a cloth, or a thread. Preferably mentioned. Note that the shape of the detection site is not limited to a line shape, and the shape of the insoluble carrier itself is not limited to a plate shape as shown in FIG.

  In the chromatographic carrier 83, a control line C for determining the end of measurement is prepared as necessary. The specific binding substance can be fixed to the test line T by, for example, a method in which the specific binding substance is directly fixed to a part of the chromatographic carrier 83 by physical or chemical binding, or the specific binding substance is latex particles. This is performed by a method in which the fine particles are physically or chemically bonded to the fine particles and trapped on a part of the chromatographic carrier 83 and immobilized.

  The labeling substance holding pad 82 is prepared, for example, by preparing a suspension containing the labeling substance, applying the suspension to an appropriate pad (for example, a glass fiber pad), and drying the suspension. is there. As a result, when the sample liquid is injected into the insoluble carrier 21, the labeling substance is allowed to flow to the region near the test line 21a while being developed together with the sample liquid. In the present invention, the labeling substance is used as an index for calculating the development rate. Further, the labeling substance may be configured to develop independently of the test substance, or may be configured to specifically bind to the test substance.

  The sample addition pad 81 is a part for spotting a sample (specimen solution or the like), and also a part for filtering insoluble matter particles or the like in the sample. In addition, the sample addition pad 81 prevents nonspecific adsorption in advance in order to prevent the test substance in the sample liquid from adsorbing nonspecifically on the material of the sample addition part and reducing the accuracy of the measurement. Sometimes processed and used.

  The absorption pad 84 is a part that absorbs and removes unreacted substances that are not insolubilized in the test line T of the chromatographic carrier 83 while the added sample is physically absorbed by the chromatographic movement. The development speed of the sample after the development front of the added sample (the boundary between the wet area and the dry area of the insoluble carrier 21) reaches the absorption pad 84 depends on the material, size, etc. of the absorbent material of the absorption pad 84. Since it is different, the speed suitable for the measurement of the test substance can be set by the selection.

  The information display unit 25 displays information related to the inspection by handwriting or sticker attachment. Examples of information relating to the examination include information relating to the patient from whom the specimen is collected (name, age, sex, etc.), information relating to the sample / reagent used in the examination (name of specimen to be examined, etc.), and the like.

(Labeling substance)
The labeling substance that can be used in the present invention is not particularly limited as long as it can detect individual particles such as fluorescent dyes, metal fine particles, and colored latex particles used in general immunochromatography. Can be used. In the present embodiment, the development speed is calculated based on the change in the position of the labeling substance represented in the image. The labeling substance may include a substance for labeling the test substance separately from the substance for obtaining the development rate. In this case, for example, a metal fine particle treated with a fluorescent substance such as a fluorescent dye as a labeling substance for obtaining the development speed and specifically bound to the test substance as a labeling substance for labeling the test substance A light-absorbing substance such as can be used. The place where the labeling substance is disposed is not limited to the case of the insoluble carrier as described above.

(Image acquisition unit)
The image acquisition unit 40 acquires an image of the test line vicinity region 21 a through the observation window 24 of the device 20. The image acquisition unit 40 corresponds to an image acquisition unit according to the present invention, and optical information in the test line vicinity region 21a is acquired based on the image. As shown in FIG. 2, the image acquisition unit 40 includes an image sensor 42 and a light source 44. When the device 20 is loaded in the measurement apparatus 1, these are arranged below the device 20 and face the observation window 24. Is configured to do. Then, the image acquisition unit 40 acquires an image of the area with the image sensor 42 while illuminating the test line vicinity area 21 a with the light source 44. Thereby, it becomes possible to grasp how the insoluble carrier 21 is developed over time based on a plurality of images. In order to save unnecessary work, the image acquisition interval is increased until the sample liquid development front can be confirmed from the observation window 24, and the image acquisition interval is obtained after the sample liquid development front can be confirmed from the observation window 24. Is preferably shortened. The image data acquired by the image acquisition unit 40 is transmitted to the memory 32. As will be described later, based on the plurality of images, the developing speed of the sample liquid, the coloration amount of the test line T, and the like are calculated.

  The image sensor 42 is configured to include, for example, a line sensor or an area sensor in which a plurality of photodiodes are arranged one-dimensionally or two-dimensionally, or an optical sensor such as a CCD, according to the luminance of received light. Produces output. The light receiving range of the image sensor 42 is, for example, a band extending in the longitudinal direction of the device 20. The light source 44 is, for example, a module that incorporates an LED, and is configured to emit white light. The light source 44 may emit monochromatic light, for example. Further, when the light source 44 includes a plurality of modules, a plurality of modules that emit monochromatic light having different wavelengths can be used. The light emitted from the light source 44 can illuminate the longitudinal direction of the device 20.

(Information reading unit)
The information reading unit 50 irradiates the information display unit 25 of the device 20 with illumination light and acquires information displayed on the information display unit 25. The method for acquiring information is not particularly limited, and the information display unit 25 can be imaged as it is or read from bar-coded information. As shown in FIG. 2, the information reading unit 50 includes an image sensor 52 and a light source 54, and when the device 20 is loaded on the measuring apparatus 1, these are arranged above the device 20 and are displayed on the information display unit 25. It is comprised so that it may oppose. Then, the information related to the inspection acquired by the information reading unit 50 and the inspection result are linked and managed. Information data acquired by the information reading unit 50 is transmitted to the memory 32. The image sensor 52 and the light source 54 are the same as the image sensor 42 and the light source 44, respectively.

(memory)
The memory 32 stores image data acquired by the image acquisition unit 40, information data acquired by the information reading unit 50, and the like. The memory 32 is configured to be able to read stored data in response to a request from the control unit 30, for example. In the present embodiment, for example, stored image data is used in the calculation processing unit 34.

(Calculation processing part)
The calculation processing unit 34 performs predetermined calculation processing based on the plurality of images acquired by the image acquisition unit 40. The calculation processing unit 34 corresponds to calculation processing means in the present invention. Specifically, the calculation processing unit 34 determines the color development amount of the test line T at a predetermined time after the arrival time when the sample liquid reaches the test line T based on the plurality of images. Further, the total passing liquid amount is calculated based on the developing speed and the cross-sectional area of the insoluble carrier, and the coloration amount at the predetermined time is normalized based on the total passing liquid amount.

The deployment speed is calculated as follows. FIG. 5 is a schematic view showing the state of the sample liquid 60 that develops in the insoluble carrier 21. In this embodiment, the insoluble carrier 21 in which the labeling substance 62 is arranged in the region upstream of the test line T is used (FIG. 5a). When the sample liquid 60 is spotted on the insoluble carrier 21, the labeling substance 62 develops in the insoluble carrier 21 with the development of the sample liquid 60 (FIGS. 5b to 5d). Then, an image of the insoluble carrier 21 (including at least the region near the test line) is acquired at a certain time t ₁ (FIG. 5b), and then an image of the insoluble carrier 21 is also acquired at a certain time t ₂ (FIG. 5c). . Here, one is a comparison of the two images while focusing on the labeling substance 62a, the distance to which the labeling substance 62a during the period from time t ₁ to t ₂ is deployed (i.e. the position of the labeling substance 62a Change). This change in the position of the labeling substance 62a is considered to correspond to the developed distance of the sample liquid 60 within that time. Therefore, the development speed of the sample liquid 60 is calculated by dividing the change in position by the time interval. The expansion rate is used as the value at time t _2. Note that if the interval for acquiring images (the interval between the times t ₁ and t _{2 in} the above) is too large, it becomes difficult to grasp the correspondence relationship between the labeling substances 62 at the times t ₁ and t _2. Is adjusted so that the correspondence between the labeling substances before and after the time can be grasped.

Also, once the deployment speed is calculated, the deployment speed does not have to be obtained at a later time. If the obtained development speed is continuously used during the measurement, the calculation process can be saved. However, since the development speed of the sample liquid 60 is not always constant during the measurement, it is preferable to calculate the development speed over time and update the value in order to perform more accurate measurement. That is, for example, as the development rate at the time _{t 3} (FIG. 5d), the image at time _{t 2} images and time _{t 3} of the insoluble carrier 21 obtained (FIG. 5c) (Figure 5d) insoluble carrier 21 obtained in It is preferable to use a value calculated by comparing. At this time, attention may be paid again to the labeling substance 62a that was noticed at the previous time, but from the viewpoint of more accurately obtaining the amount of liquid passing through the test line T during this time, When a close labeling substance exists, it is preferable to pay attention to the labeling substance. That is, it is preferable to use a change in the position of the labeling substance existing in the vicinity of the detection site before and after the change as the change in the position of the labeling substance. In FIG. 5, for example, rather than focusing on the labeling substance 62a that has already passed the test line T at time t _2, focusing on the labeling substance 62b which passes through the test line T from the time t ₂ to time t ₃ It is preferable.

  In the above, the development speed is calculated by paying attention to one certain labeling substance 62, but it may be calculated by paying attention to a plurality of labeling substances 62 and averaging the values obtained for each labeling substance 62. .

  The time when the sample liquid 60 reaches the test line T is the time when the development front contacts the test line T. As the time, for example, a time at which an image in which contact of the development front with the test line T is first confirmed by image analysis is acquired, or a time calculated based on the development speed at that time is used. Can be.

  The coloration amount of the test line T is calculated on the basis of the optical density and chromaticity, for example, by obtaining the optical density and chromaticity of the pixel region corresponding to the test line T in the image by image analysis. For the cross-sectional area of the insoluble carrier 21, for example, a value measured in advance is set in the measuring device 1.

The total amount of liquid passing through at a predetermined time means the amount of sample liquid that has actually passed through the test line T during the period from the arrival time to the predetermined time. For example, when the arrival time is t _α , the total amount of liquid passing through at time t _N is [average development speed from arrival time t _α to time t _N ] × [cross-sectional area of the insoluble carrier] × [t _N − It can be calculated by the calculation formula 1 of t _α ] or the calculation formula 2 of Σ [development speed at time t _n ] × [cross-sectional area of the insoluble carrier] × [t _n −t _n−1 ]. Here, Σ means that the sum of α + 1 to N is taken for _n , and [development speed at time t _n ] × [cross-sectional area of insoluble carrier] × [t _n −t _n−1 ] is time t _n. It means a partial flow-through amount of from _-1 to t _n.

  The normalization of the coloring amount is performed, for example, by dividing the measured value of the coloring amount by the total passing liquid amount.

  Hereinafter, the steps of the measurement method according to the present embodiment will be described with reference to FIG. FIG. 6 is a schematic diagram showing a workflow of the process of the measurement method in the present embodiment.

First, the user loads the device 20 into which the sample liquid has been injected from the device loading unit 14 into the measurement apparatus 1 (STEP 1). The measuring apparatus 1 conveys the device 20 inside, and arrange | positions the device 20 in the predetermined position which can be measured. At this time, the information reading unit 50 reads predetermined information. The measuring device 1 resets the time t _n (n = 0) when it reaches a state where the measurement can be started.

Then, after the time has been reset, the first image is acquired at time _{t 1} (STEP2). At this point in time, the development front usually does not reach the test line T, so that it is not particularly necessary to calculate the development speed and the coloration degree. Thereafter, image acquisition at time t _n is performed with time (that is, n is incremented) (STEP 3). At this time, each time an image is acquired, it is determined based on the image whether the sample liquid has reached the test line T (STEP 4). As a result of the determination, if it is determined that the sample liquid has not reached the test line T, the process is repeated from STEP3. If it is determined that the sample liquid has reached the test line T, the current time t _{n is determined.} Is stored as t _α (STEP 5).

Similarly, after the time t _α elapses, image acquisition is performed with time (STEP 6). However, after the time t _{α has} elapsed, every time an image is acquired, the development speed (STEP 7), the total passing liquid amount (STEP 8), and the coloration amount (STEP 9) at the time t _n are calculated based on the image. . In this embodiment, deployment speed at time t _n is calculated based on a change in the position of the labeling substance which is grasped by comparing the image in the image and the time t _n at time t _n-1, as described above . The total effluent volume at time t _n, from the arrival time t _alpha of the stored arrival time t _alpha Toko based on expansion rate of the average to the time t _n, are calculated for example by the above formula 1. When the total passing liquid amount and the coloring amount at time t _n are calculated, the coloring amount is normalized by dividing the coloring amount by the total passing liquid amount, and the data is stored in the memory 32, for example. (STEP 10).

When the normalization of the coloring amount at time t _n is completed, it is determined whether or not the measurement is completed (STEP 11). Whether or not the measurement is completed is determined based on the coloration amount of the control line C, for example. As a result of this determination, if it is determined that the measurement is not completed, the process is repeated from STEP6.

  As described above, according to the chromatograph measurement method and the measurement apparatus according to the present invention, the total amount of liquid passing through from the arrival time to a predetermined time is calculated, and the presentation at the predetermined time is based on the total amount of liquid passing through. The amount of color can be normalized. That is, the measured value of the coloration amount is corrected by the total passing liquid amount. As a result, in chromatographic measurement, even if the total amount of liquid passing through each measurement cannot be made uniform, measurement with higher quantitativeness becomes possible.

<Design changes>
In the description of the embodiment, the step of calculating the total passing liquid amount at the time t _n by the calculation formula 1 has been described. However, for example, when calculating the total passing liquid amount at the time t _n by the calculation formula 2, the step Is as follows.

  FIG. 7 is a schematic diagram illustrating a workflow of a measurement method process in the design change of the embodiment. Since this step is the same as the embodiment up to STEP 4, the description thereof is omitted.

If it is determined in STEP 4 that the sample liquid has reached the test line T, image acquisition is performed over time in the same manner (STEP 5). It is not particularly necessary to store the arrival time t _alpha in this process. At time t _alpha elapsed after deployment speed at time _{t n} on the basis of the image every time the image is acquired (STEP6), passing liquid amount from time _{t n-1} to time _{t n} (STEP7), the time t A total passing liquid amount at _n (STEP 8) and a coloration amount at time t _n (STEP 9) are calculated. The amount of liquid passing through from time t _n-1 to time t _n is calculated by [development speed at time t _n ] × [cross-sectional area of insoluble carrier] × [t _n −t _n−1 ], and at time t _n the total permeate volume is calculated by adding the flow-through amount from time t _n-1 to the total flow-through volume to the time t _n at time t _n-1. The total effluent volume at arrival time t _alpha as the initial value is zero. When the total passing liquid amount and the coloring amount at time t _n are calculated, the coloring amount is normalized by dividing the coloring amount by the total passing liquid amount, and the data is stored in the memory 32, for example. (STEP 10).

When the normalization of the coloring amount at time t _n is completed, it is determined whether or not the measurement is completed (STEP 11). As a result of this determination, if it is determined that the measurement is not completed, the process is repeated from STEP5.

In the description of the embodiment, the development speed is calculated based on the change in the position of the labeling substance, but the present invention is not limited to this. For example, as shown in FIG. 8, the development speed can also be calculated based on the change in the position of the development front 64 on the insoluble carrier 21. Specifically, by dividing the amount of change in the position of the development front 64 obtained by comparing the image at time t ₁ (FIG. 8a) and the image at time t ₂ (FIG. 8b) by the time interval, The deployment speed is calculated.

DESCRIPTION OF SYMBOLS 1 Chromatographic measurement apparatus 10 Case 11 Screen display part 12 Menu operation part 13 Power switch 14 Device loading part 20 Immunochromatography device 21 Insoluble carrier 21a Test line vicinity area 22 Device case 23 Solution inlet 24 Observation window 25 Information display Unit 30 Control unit 32 Memory 34 Calculation processing unit 40 Image acquisition unit 50 Information reading unit 60 Sample liquid 62 Labeling substance 64 Development front C Control line T Test line

Claims (9)

In the chromatographic measurement method for detecting the test substance using an insoluble carrier having a detection site capable of specifically fixing the test substance contained in the sample liquid,
Developing the sample solution in the insoluble carrier;
Based on a plurality of images of the insoluble carrier acquired over time by the image acquisition means, the sample during a period from the arrival time at which the sample liquid reaches the detection site to a predetermined time after the arrival time Calculating the development speed of the liquid a plurality of times over time, and calculating the coloration amount of the detection site at the predetermined time ;
Based on the calculated plurality of development speeds and the cross-sectional area of the insoluble carrier, the amount of the sample liquid that has actually passed through the detection site between the arrival time and the predetermined time is calculated. ,
A chromatographic measurement method characterized by normalizing the coloration amount at the predetermined time based on the calculated amount of the sample liquid. The labeling substance is developed in the insoluble carrier together with the sample liquid,
The chromatographic measurement method according to claim 1, wherein the development speed is calculated based on a change in position of the labeling substance represented in the plurality of images.   The chromatographic measurement method according to claim 2, wherein the labeling substance is disposed in a region upstream of the detection site.   The chromatographic measurement method according to claim 2, wherein the labeling substance is previously mixed in the sample liquid.   The chromatographic measurement method according to claim 1, wherein the development speed is calculated based on a change in a position of a development front of the sample liquid. In the chromatographic measurement apparatus for detecting the test substance using an insoluble carrier having a detection site capable of specifically fixing the test substance contained in the sample liquid,
The insoluble carrier;
Image acquisition means for acquiring a plurality of images of the insoluble carrier over time;
Based on the plurality of images, the development speed of the sample liquid is calculated a plurality of times over time during a period from the arrival time when the sample liquid reaches the detection site to a predetermined time after the arrival time. And the amount of coloration of the detection part at the predetermined time , and based on the calculated plurality of development speeds and the cross-sectional area of the insoluble carrier, the time period from the arrival time to the predetermined time is calculated. A calculation processing unit that calculates an amount of the sample liquid that has actually passed through the detection site and normalizes the coloration amount at the predetermined time based on the calculated amount of the sample liquid. A chromatograph measuring device characterized by the above. The calculation processing means is a labeling substance that develops in the insoluble carrier together with the sample solution, and calculates the development speed based on a change in the position of the labeling substance represented in the plurality of images. The chromatograph measuring apparatus according to claim 6 . The chromatograph measuring apparatus according to claim 7 , comprising the labeling substance disposed in a region upstream of the detection site. The chromatograph measuring apparatus according to claim 6 , wherein the calculation processing means calculates the development speed based on a change in the position of the development front of the sample liquid.

Images